钙通道阻滞剂维拉帕米抑制肝星状细胞活化和功能

Verapamil inhibits the activation and function of hepatic stellate cells

探讨维拉帕米(Ver)对人肝星状细胞系(HSC)-LX2的活化及分泌转化生长因子(TGF-β)的抑制作用。方法体外用内皮素-1(endothelin,ET-1)刺激HSC-LX2活化建立培养体系,设对照、ET-1和ET-1+Ver 3组,对照组仅加入培养基,ET-1组加入ET-1和培养基,ET-1+Ver组加入ET-1、Ver和培养基,分别在(0、12、24、48、72 h)观察HSC分沁α-平滑肌肌动蛋白(α-SMA)和细胞因子的能力,采用细胞爬片和免疫组化技术鉴定活化HSC的细胞形态;流式细胞术分析HSC表达α-SMA水平,ELISA测定不同组HSC分泌TGF-β的浓度。结果与对照组比较,ET-1组HSC活化和分泌TGF-β的能力较强(P<0.05);与ET-1组比较,ET-1+Ver组HSC活化和分泌TGF-β能力较弱(P<0.05)。结论 Ver体外实验中能抑制HSC的增殖和活化,具有潜在的抗纤维化作用。

Objective To investigate the effect of Verapamil on the regulation of proliferation and function of human hepatic stellate cell lines（HSC）,and illuminate its further pharmacological role.Methods Activation of HSC culture system stimulated by ET-1 was established in vitro,which was divided into three groups： control group,ET-1 group and ET-1＋Verapamil group.The control group was added with medium only.The ET-1 group was added with medium and ET-1.The ET-1＋Verapamil group was added with medium,ET-1 and Verapamil.Slide of crawling cell was adopted to identify morphology of HSC.Flow cytometry was employed to detect the activation of HSC by analyzing the level of α-SMA.ELISA was used to determine the concentration of TGF-β.Results Compared with control group,the expression of α-SMA and TGF-β of HSC in vitro significantly increased in ET-1group（P0.05）.However,the expression of α-SMA and TGF-β of HSC significantly decreased in ET-1＋Verapamil group compared with ET-1 group（P0.05）.Conclusion Verapamil can inhibit the proliferation and activation of HSC.Verapamil can suppress the occurrence of liver fibrosis.